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蛋白质糖基化(glycosylation)是最常见和最重要的翻译后修饰之一.大规模N-连接糖基化位点鉴定是糖蛋白质组学研究的重要组成部分,而N-连接糖肽富集是高通量N-连接糖基化位点鉴定的关键步骤.凝集素富集法和酰肼化学法是目前被广泛应用的N-连接糖肽富集技术,有报道认为两种方法具有很强的互补性,联合使用能提高糖基化位点的鉴定数目.本文以Hep G2细胞系为模型,系统比较了这两种方法的富集效率和糖蛋白鉴定数目.结果表明,虽然酰肼法的糖肽富集效率为76.6%,远高于凝集素法的54.6%,但是凝集素法却能鉴定到825个糖蛋白和1 959个N-连接糖基化位点,显著多于酰肼法富集到的522个糖蛋白和1 014个糖基化位点.并且,两种方法并未显示出显著的互补性,仅28个糖蛋白和80个糖基化位点未在凝集素法中鉴定到.
Glycosylation of proteins is one of the most common and important post-translational modifications. Massive N-linked glycosylation site identification is an important part of the glycoproteomics study, whereas N-linked glycopeptide enrichment Is a key step in the identification of high-throughput N-linked glycosylation sites.Lectin-rich and hydrazide chemistries are the most widely used N-linked glycopeptide enrichment technologies and it has been reported that both methods have very good Strong complementarity, combined use can increase the number of glycosylation sites identified.In this paper, Hep G2 cell line as a model, the system compared the efficiency of these two methods and glycoprotein identification number.The results show that although the hydrazide The glycopeptide enrichment efficiency of the method was 76.6%, much higher than that of the lectin method (54.6%), but the lectin method could identify 825 glycoproteins and 1 959 N-linked glycosylation sites, which were significantly more than that of the acyl Hydrazine method enriched in 522 glycoproteins and 1 014 glycosylation sites and the two methods did not show significant complementarity with only 28 glycoproteins and 80 glycosylation sites not in the aggregation Supra Footwear identified in the method.